A Comprehensive Review on HLA and its Detections by Polymerase Chain Reaction Technique

Author(s)

Chatterjee, A., Sinha, S. K., Mukherjee, G.

Author's Affiliation

Abstract

The HLA or histocompatibility antigens play predominant role in acceptance or rejection of transplanted organs and in the regulation of immune response as well as in susceptibility or resistance to a large number of diseases. This system is highly polymorphic. Perfect HLA testing is also essential for anthropological studies having different racial groups, for blood component therapy as well as for research application in the development of MHC based vaccines. Now-a-days molecular (PCR) based HLA technique is recommended for HLA typing. Sequence-Specific Primer PCR (PCR-SSP) technique is based on the specificity of the primers in which a 3’ single-base mismatch inhibits the priming of a non-specific reaction. Among the different molecular method usually PCR-SSP is done as it is comparatively easier and cost effective. Sequence Specific Oligonucleotide Probe Hybridization (PCR-SSOP) is more amenable to high-throughput HLA typing than PCR-SSP. In Reverse Line Strip (RLS) or PCR-SSO reverse assay multiple oligonucleotide probes specific for alleles of interest are immobilized on a single membrane which is hybridized with amplified DNA and the detection of HLA alleles is possible by colour development using a specific substrate. Luminex technology is another variation of PCR-SSO reverse assay where membrane is replaced with 100 different colours of polystyrene microbead. Reference strand–mediated conformational analysis (RSCA) is a conformational method for high-resolution typing class I and II genes using native polyacrylamide gels. Genomic DNA sequencing–based typing (SBT) methodology utilizes locus- or group-specific amplification followed by cycle-sequencing the desired number of exons on both strands.

Keywords

PCR-SSP, PCR-SSOP, RLS, RSCA, Luminex Technology

Cite This Article

Chatterjee, A., Sinha, S. K., & Mukherjee, G. A Comprehensive Review on HLA and its Detections by Polymerase Chain Reaction Technique. International Journal for Pharmaceutical Research Scholars (IJPRS), 3(2), 340-346.